PIB ATPases are metal cation pumps that transport metals across membranes. These proteins possess N- and C-terminal cytoplasmic extensions that contain Cys- and His-rich high affinity metal binding domains, which may be involved in metal sensing, metal ion selectivity and/or in regulation of the pump activity. The PIB ATPase HMA4 (Heavy Metal ATPase 4) plays a central role in metal homeostasis in Arabidopsis thaliana and has a key function in zinc and cadmium hypertolerance and hyperaccumulation in the extremophile plant species Arabidopsis halleri. <br />Here, we examined the function and structure of the N-terminal cytoplasmic metal-binding domain of HMA4. We mutagenized a conserved CCTSE metal-binding motif in the domain and assessed the impact of the mutations on protein function and localization in planta, on metal-binding properties in vitro and on protein structure by Nuclear Magnetic Resonance spectroscopy. <br />The two Cys residues of the motif are essential for the function, but not for localization, of HMA4 in planta, whereas the Glu residue is important but not essential. These residues also determine zinc coordination and affinity. Zinc binding to the N-terminal domain is thus crucial for HMA4 protein function, whereas it is not required to maintain the protein structure. <br />Altogether, combining in vivo and in vitro approaches in our study provides insights towards the molecular understanding of metal transport and specificity of metal P-type ATPases. [less ▲]

in Proceedings of the National Academy of Sciences (2016), 113(13), 3557-62

Many experiments have shown that local biodiversity loss impairs the ability of ecosystems to maintain multiple ecosystem functions at high levels (multifunctionality). In contrast, the role of ... [more ▼]

Many experiments have shown that local biodiversity loss impairs the ability of ecosystems to maintain multiple ecosystem functions at high levels (multifunctionality). In contrast, the role of biodiversity in driving ecosystem multifunctionality at landscape scales remains unresolved. We used a comprehensive pan-European dataset, including 16 ecosystem functions measured in 209 forest plots across six European countries, and performed simulations to investigate how local plot-scale richness of tree species (α-diversity) and their turnover between plots (β-diversity) are related to landscape-scale multifunctionality. After accounting for variation in environmental conditions, we found that relationships between α-diversity and landscape-scale multifunctionality varied from positive to negative depending on the multifunctionality metric used. In contrast, when significant, relationships between β-diversity and landscape-scale multifunctionality were always positive, because a high spatial turnover in species composition was closely related to a high spatial turnover in functions that were supported at high levels. Our findings have major implications for forest management and indicate that biotic homogenization can have previously unrecognized and negative consequences for large-scale ecosystem multifunctionality. [less ▲]

Preserving ecosystem functions of soil organic matter (SOM) in soils is a key challenge. The need for an efficient diagnosis of SOM state in agricultural soils is a priority in order to facilitate the ... [more ▼]

Preserving ecosystem functions of soil organic matter (SOM) in soils is a key challenge. The need for an efficient diagnosis of SOM state in agricultural soils is a priority in order to facilitate the detection of changes in soil qualityas a result of changes in management practices. The nature of SOM is complex and cannot readily be monitored due to the heterogeneity of its components. Assessment of the SOM level dynamics, typically characterized as the bulk soil organic carbon (SOC), can be refined by taking into account carbon pools with different turnover rates and stability. Fractionating bulk SOC in meaningful soil organic fractions helps to better diagnose SOC status. By separating carbon associated with clay and fine silt particles (stable carbon with slow turnover rate) and carbon nonassociated with this fraction (labile and intermediate carbon with higher turnover rates), effects of management can be detected more efficiently at different spatial and temporal scales. Until now, most work on SOC fractionation has focused on small spatial scales along management or time gradients. The present case study focuses on SOC fractionation applied in order to refine the interpretation of organic matter turnover and SOC sequestration for regional units in Wallonia with comparable climate, management and, to a certain extent, soil conditions. In each unit, random samples from specific land uses are analyzed in order to assess the Normal Operative Ranges (NOR) of SOC fraction contents for each unit and land use combination. Thus, SOC levels of the different fractions of a specific field in a given unit can be compared to its corresponding NOR. It will help to better diagnose agricultural soil quality in terms of organic carbon compared to a bulk SOC diagnosis. [less ▲]

As many other countries, Belgium complies with Annex I of the United Nations Framework Convention on Climate Change (UNFCCC). Belgium thus reports its annual greenhouse gas emissions in its national ... [more ▼]

As many other countries, Belgium complies with Annex I of the United Nations Framework Convention on Climate Change (UNFCCC). Belgium thus reports its annual greenhouse gas emissions in its national inventory report (NIR), with a distinction between emissions/sequestration in cropland and grassland (EU decision 529/2013). The CO2 fluxes are then based on changes in SOC stocks computed for each of these two types of landuse. These stocks are specified for each of the agricultural regions which correspond to areas with similar agricultural practices (rotations and/or livestock) and yield potentials. For Southern Belgium (Wallonia) consisting of ten agricultural regions, the Soil Monitoring Network (SMN) ‘CARBOSOL’ has been developed this last decade to survey the state of agricultural soils by quantifying SOC stocks and their evolution in a reasonable number of locations complying with the time and funds allocated. Unfortunately, the 592 points of the CARBOSOL network do not allow a representative and a sound estimation of SOC stocks and its uncertainties for the 20 possible combinations of land use/agricultural regions. Moreover, the SMN CARBIOSOL is based on a legacy database following a convenience scheme sampling strategy rather than a statistical scheme defined by design-based or model-based strategies. Here, we aim to both quantify SOC budgets (i.e. How much?) and spatialize SOC stocks (i.e. Where?) at regional scale (Southern Belgium) based on data from the SMN described above. To this end, we developed a computation procedure based on Digital Soil Mapping techniques and stochastic simulations (Monte-Carlo) allowing the estimation of multiple (10,000) independent spatialized datasets. This procedure accounts for the uncertainties associated to estimations of both i) SOC stock at the pixelscale and ii) parameters of the models. Based on these 10,000 individual realizations of the spatial model, mean SOC stocks and confidence intervals can be then computed at the pixel scale, for selected sub-areas (i.e. the 20 landuse/agricultural region combinations) and for the entire study area. [less ▲]

Earthworms have a significant influence on the structure, composition and functioning of forest ecosystems, but in spite of their role as ecosystem engineers, little is known on the factors controlling ... [more ▼]

Earthworms have a significant influence on the structure, composition and functioning of forest ecosystems, but in spite of their role as ecosystem engineers, little is known on the factors controlling their distribution across European forests. Optimised sampling techniques, as well as more advanced statistical tools and geographical information systems have facilitated studies at the landscape scale. But these, and even larger-scale studies, are scarce due to data limitations, taxonomic inconsistencies and practical issues in linking existing databases. In this continental-scale field-based study we used boosted regression tree modelling to identify and evaluate the relative importance of environmental factors explaining earthworm incidence (presence/absence) and abundance (density and biomass) in European forests. To parameterise our models earthworms were sampled in six forest landscapes along a latitudinal gradient from the boreal north to the Mediterranean south in spring or autumn of 2012, together with several environmental variables. Earthworms were sampled using a combined method of mustard extraction and hand sorting of litter and a soil monolith, after which they were weighed and identified to functional group (epigeic, endogeic and anecic). We found that litter- and soil-related variables best explained earthworm incidence and biomass in European forests, leaving only a minor role to climaterelated variables. Among the litter related variables, understory vegetation played an important role in explaining earthworm incidence and abundance. The relative importance of explanatory variables differed between models for incidence, density and biomass and between earthworm functional groups. Our results suggested that threshold values for soil C:N ratio, forest floor pH and understory plant biomass and plant nutrient concentrations have to be attained before earthworms can occur. Beyond these threshold values, variables like soil C:N ratio, tree litter C:P ratio and forest floor mass further explain earthworm biomass. Mechanisms behind these observations are discussed in the light of future earthworm distribution modelling at continental scale. [less ▲]

There is considerable evidence that biodiversity promotes multiple ecosystem functions (multifunctionality), thus ensuring the delivery of ecosystem services important for human well-being. However, the ... [more ▼]

There is considerable evidence that biodiversity promotes multiple ecosystem functions (multifunctionality), thus ensuring the delivery of ecosystem services important for human well-being. However, the mechanisms underlying this relationship are poorly understood, especially in natural ecosystems. We develop a novel approach to partition biodiversity effects on multifunctionality into three mechanisms and apply this to European forest data. We show that throughout Europe, tree diversity is positively related with multifunctionality when moderate levels of functioning are required, but negatively when very high function levels are desired. For two well-known mechanisms, 'complementarity' and 'selection', we detect only minor effects on multifunctionality. Instead a third, so far overlooked mechanism, the 'jack-of-all-trades' effect, caused by the averaging of individual species effects on function, drives observed patterns. Simulations demonstrate that jack-of-all-trades effects occur whenever species effects on different functions are not perfectly correlated, meaning they may contribute to diversity-multifunctionality relationships in many of the world's ecosystems. [less ▲]

Tree species influence biogeochemical cycling through element deposition (throughfall, litterfall), root decomposition and exudates, and through their influence on the microbial activities in the soil ... [more ▼]

Tree species influence biogeochemical cycling through element deposition (throughfall, litterfall), root decomposition and exudates, and through their influence on the microbial activities in the soil. Yet, the effect of mixing tree species on soil functioning is unclear, in particular concerning the microbial diversity and activity in soils. Here we synthesize results from the Exploratory Platform of the FunDivEUROPE project (http://www.fundiveurope.eu/). This network of 209 comparative plots covering tree diversity levels of 1 to 5 species was established in existing mature forests in 6 European regions. These six focal regions represent a gradient of major European forest types from boreal to Mediterranean forests. The aims of this study were to determine the soil microbial biomass and metabolic diversity of soil bacteria for these 6 European forest regions, presenting each a tree species richness gradient and to analyse the impact of tree species richness and the role of other controlling factors. We analysed the relation between tree species diversity, the proportion of coniferous tree species and soil factors (pH, soil organic carbon, water soluble carbon and nitrogen) and the carbon substrate utilisation pattern of soil bacteria (BIOLOG Ecoplate), soil microbial biomass (fumigation-extraction), hot water carbon and nitrogen in the forest floor and the upper mineral soil horizon (linear mixed models, GLM for multivariate abundance data, discriminant correspondence analysis). Mean values of microbial biomass carbon ranged from 3264 (Italy) to 8717 (Finland) mg kg-1 in the forest floor and from 465 (Italy) to 3748 (Finland) mg kg-1 in the mineral soil. Statistical models predicted microbial biomass to increase in both soil layers by 7-8% with each step increase in tree diversity. Increased proportion of conifers was linked to a decrease in the number of carbon substrates used by soil bacteria. The types of carbon sources used were dependent on region, proportion of conifers, soil pH and water-soluble carbon and nitrogen. [less ▲]

In Arabidopsis thaliana, FRD3 (Ferric Chelate Reductase Defective 3) plays a central role in metal homeostasis. FRD3 is among a set of metal homeostasis genes that are constitutively highly expressed in ... [more ▼]

In Arabidopsis thaliana, FRD3 (Ferric Chelate Reductase Defective 3) plays a central role in metal homeostasis. FRD3 is among a set of metal homeostasis genes that are constitutively highly expressed in roots and shoots of Arabidopsis halleri, a zinc hyperaccumulating and hypertolerant species. Here, we examined the regulation of FRD3 by zinc in both species to shed light on the evolutionary processes underlying the evolution of hyperaccumulation in A. halleri. We combined gene expression studies with the use of GUS and GFP reporter constructs to compare the expression profile, transcriptional and post-transcriptional regulation of FRD3 in both species. The AtFRD3 and AhFRD3 genes display a conserved expression profile. In A. thaliana, alternative transcription initiation sites from two promoters determine transcript variants which are differentially regulated by zinc supply in roots and shoots to favour the most highly translated variant under zinc excess conditions. In A. halleri, a single transcript variant with higher transcript stability and enhanced translation has been maintained. The FRD3 gene thus undergoes complex transcriptional and post-transcriptional regulation in Arabidopsis relatives. Our study reveals that a diverse set of mechanisms underlie increased gene dosage in the A. halleri lineage and illustrates how an environmental challenge can alter gene regulation. [less ▲]

Drier condition during the growing season have been predicted in the future. It has been suggested that diverse forest could maintain productivity and provide better ecosystem services under stress ... [more ▼]

Drier condition during the growing season have been predicted in the future. It has been suggested that diverse forest could maintain productivity and provide better ecosystem services under stress condition such as drought. However, those studies focused mainly on mature forest and little known about young forest. Oak and beech are the important species in European forestry, and may face a strong challenge in the future. Drought effects on young ( ̴5yr) oak and beech saplings in monoculture and mixed with other species are not known. Moreover, single studies evaluating both above- and below-ground ecosystem response to drought are scarce. A two-year manipulative field experiment has been planned to answer the following questions. (i) Can species mixtures improve oak and beech sapling performances under drought conditions? (ii) What are the mechanisms underlying ecosystem functioning and sapling performance in mixed species stands subjected to drought? A 3m × 3m rainout shelter will be placed only in growing season in Zedelgem sites of FORBIO experimental platform (http://www.treedivbelgium.ugent.be/pl_forbio.html). Tree diversity vary from 1 to 4 species and about 50% of precipitation will be taken off. Both aboveground sapling performance and belowground microbial properties and biogeochemical processes will be investigated. We will present the design of the experimental tree species diversity plantation of Zedelgem site, the setting of the drought experiment and planned analysis [less ▲]